Application of STING inhibitors in the preparation of drugs for treating or preventing post-cardiac arrest syndrome

Abstract

The present invention relates to the field of biomedicine, and more specifically, to the use of STING inhibitors in the preparation of medicaments for treating or preventing post-cardiac arrest syndrome. The present invention proposes that STING inhibitors, through their interventional effects, inhibit STING-mediated type I interferon activation, thereby alleviating brain damage in patients with post-cardiac arrest syndrome and improving their survival rate and prognosis.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of a STING inhibitor in the preparation of a drug for treating or preventing post-cardiac arrest syndrome. Background Art

[0002] Millions of cardiac arrests (CAs) occur worldwide annually. In the United States, for example, the incidence of out-of-hospital cardiac arrest (OHCA) is approximately 140 per 100,000 patients, with a survival rate of approximately 10%. In contrast, approximately 209,000 in-hospital cardiac arrests (IHCA) occur annually, with a survival rate of only 24%, of which 14% develop significant neurological impairment. In my country, the incidence of CA is 40.7 per 100,000 patients, with only 1% of patients surviving with a good neurological outcome. Therefore, further improving the survival rate and neurological outcomes of CA patients remains a key research priority and challenge. Post-cardiac arrest syndrome (PAS) is the leading cause of mortality in patients after resuscitation from cardiac arrest and a major cause of long-term disability in acute survivors. Despite current measures such as targeted temperature management and corticosteroids to improve neurological outcomes in patients after cardiopulmonary resuscitation, up to 60% of patients still experience neurological deficits. Excessive release of immune and inflammatory mediators is a key factor in the organ damage caused by PAS. Therefore, it is crucial to find targets that regulate immune inflammatory mediators to alleviate organ damage in post-cardiac arrest syndrome.

[0003] The cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway is crucial for sensing abnormal DNA accumulation in the cytoplasm and is a major pathway mediating DNA-mediated immune responses in vivo. When exogenous DNA from bacteria, viruses, and tumor cells, or autologous DNA from mitochondrial damage or apoptosis, accumulates in the cytoplasm, it is recognized and bound by cGAS to generate the second messenger cyclic dinucleotide 2',3'-cGAMP. 2',3'-cGAMP then transmits the signal to the downstream STING, inducing the translocation of transcription factors such as IRF3 and NF-κB into the cell nucleus, thereby inducing the expression of inflammatory cytokines such as type I interferons, ultimately initiating an immune response. As a central immune molecule in numerous signaling pathways, STING is normally in an autoinhibitory state as a dimer. Upon stimulation by upstream signals, it undergoes a conformational change, leading to its activation. This in turn triggers the activation and production of a series of downstream immune cells and inflammatory factors, promoting the onset of inflammatory responses and the corresponding clinical symptoms.

[0004] Currently, the main treatments for cardiac arrest are epinephrine, lidocaine, sodium bicarbonate, and atropine, which are primarily used to increase circulatory perfusion and improve ventilation. Few drugs have been reported to protect against organ damage in post-cardiac arrest syndrome.

[0005] H151 is a potent, selective, and covalent STING inhibitor with significant anti-inflammatory activity both in vitro and in vivo. It can suppress excessive immune responses, alleviate diseases such as sepsis, and protect the kidneys and liver. However, whether the STING signaling pathway is involved in the pathological process of post-cardiac arrest syndrome and the specific mechanism have not been reported, and whether STING inhibitors can be used to treat post-cardiac arrest syndrome has not been studied.

[0006] In view of this, the present invention is proposed. Summary of the Invention

[0007] In order to address the technical defects of the existing technology and find a drug for reducing organ damage in post-cardiac arrest syndrome, the present invention provides the use of a STING inhibitor in the preparation of a drug for treating or preventing post-cardiac arrest syndrome.

[0008] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0009] One aspect of the present invention relates to the use of a STING inhibitor in the preparation of a medicament for treating and / or preventing post-cardiac arrest syndrome.

[0010] STING inhibitors are a drug intervention method targeting the STING pathway. STING is the main signaling molecule in the intracellular DNA sensing pathway and plays an important role in the innate immune response.

[0011] Post-cardiac arrest syndrome refers to a series of physical and psychological symptoms caused by the interruption or reduction of cardiac blood pumping during cardiac arrest, even though the autonomous heartbeat is restored after rescue treatment.

[0012] The present invention proposes to alleviate brain damage in patients with post-cardiac arrest syndrome and improve their survival rate and prognosis through the intervention of STING inhibitors to inhibit STING-mediated type I interferon activation.

[0013] The present invention found that knocking out the STING gene or using a STING inhibitor can significantly reduce brain tissue damage in patients with post-cardiac arrest syndrome by regulating the STING-I type interferon immune-inflammatory signaling pathway, improve patients' spatial memory function and motor function, and significantly increase the overall survival time and survival rate of patients with post-cardiac arrest syndrome.

[0014] Based on the same efficacy of STING inhibitors in treating post-cardiac arrest syndrome, STING inhibitors also have a certain effect in preventing post-cardiac arrest syndrome. For some patients who are prone to cardiac arrest, early intervention with STING inhibitors can reduce the probability and intensity of post-cardiac arrest syndrome.

[0015] The present invention does not specifically limit the source of the STING inhibitor, and any STING inhibitor conventionally purchased by those skilled in the art can be used to implement the technical solution of the present invention.

[0016] Furthermore, the STING inhibitor includes but is not limited to at least one of H-151, SN-001, SN-011 or AK59.

[0017] Furthermore, the structure of H-151 is as follows:

[0018]

[0019] The present invention selects H-151 among the STING inhibitors, which has a significant effect in treating post-cardiac arrest syndrome. It can alleviate brain damage in patients with post-cardiac arrest syndrome by inhibiting the activation of type I interferon mediated by STING, thereby improving the survival rate and prognosis of patients.

[0020] The present invention does not specifically limit the source of H-151, and any H-151 conventionally purchased by those skilled in the art can be used to implement the technical solution of the present invention.

[0021] Furthermore, the STING inhibitor can regulate the STING-mediated type I interferon immune inflammatory signaling pathway.

[0022] Furthermore, the application includes any one or more of the following:

[0023] (1) Alleviate brain damage caused by post-cardiac arrest syndrome;

[0024] (2) Improve spatial memory function and / or motor function after post-cardiac arrest syndrome;

[0025] (3) Improve the survival period and survival rate after post-cardiac arrest syndrome.

[0026] Another aspect of the present invention also relates to a drug for treating and / or preventing post-cardiac arrest syndrome, the active ingredient of which includes: a STING inhibitor.

[0027] The drug for treating and / or preventing post-cardiac arrest syndrome contains a STING inhibitor as an active ingredient. The STING inhibitor can inhibit the activation of type I interferon mediated by STING, thereby alleviating brain damage in patients with post-cardiac arrest syndrome and improving the survival rate and prognosis of patients.

[0028] Furthermore, the active ingredients of the drug for treating and / or preventing post-cardiac arrest syndrome include but are not limited to: at least one of H-151, SN-001, SN-011 or AK59.

[0029] Furthermore, the drug for treating and / or preventing post-cardiac arrest syndrome further comprises: a pharmaceutically acceptable carrier and / or a pharmaceutical excipient.

[0030] The present invention has no special limitation on pharmaceutically acceptable carriers and pharmaceutical excipients, and the technical solution of the present invention can be implemented by using conventional carriers and excipients in the art.

[0031] Furthermore, the dosage forms of the drug for treating and / or preventing post-cardiac arrest syndrome include: oral preparations and / or injection preparations.

[0032] Furthermore, the oral preparation includes at least one of: tablets, capsules, oral liquids, granules, suspensions, dropping pills, sustained-release preparations, controlled-release preparations or targeted preparations.

[0033] Furthermore, the injection preparation includes: water injection and / or powder injection.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] (1) The present invention applies STING inhibitors to the preparation of drugs for treating and / or preventing post-cardiac arrest syndrome. Through the intervention of STING inhibitors, STING-mediated type I interferon activation is inhibited, thereby significantly reducing brain tissue damage in patients with post-cardiac arrest syndrome, improving patients' spatial memory function and motor function, and significantly improving the overall survival time and survival rate of patients with post-cardiac arrest syndrome.

[0036] (2) The drug for treating and / or preventing post-cardiac arrest syndrome provided by the present invention contains a STING inhibitor as an active ingredient. The STING inhibitor can inhibit the activation of type I interferon mediated by STING, thereby alleviating brain damage in patients with post-cardiac arrest syndrome and improving the survival rate and prognosis of patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0038] Figure 1 The 7-day survival of STING knockout mice and wild-type mice after cardiac arrest and cardiopulmonary resuscitation ( Figure 1 A), neurological function scores at 3, 7, and 14 days ( Figure 1 B) and representative histological HE staining images of brain tissue ( Figure 1 C);

[0039] Figure 2 The Sham group, CPR-WT group, and CPR-STING group provided in the embodiment of the present invention - / - The changes in the phosphorylation levels of IRF3 and TBK1, key proteins in the STING downstream signaling pathway, in the mice of the cardiac arrest group after cardiopulmonary resuscitation were observed. The upregulation of the phosphorylation levels of IRF3 and TBK1 indicated the activation of the STING signaling pathway.

[0040] Figure 3 The RT-qPCR assay provided in the embodiment of the present invention detected IFN-β, IL-1β, IL-6, TNF-α, CXCL10 and CCL5 in the Sham group, CPR-WT group, and CPR-STING group. - / - Expression of IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 in brain tissue of mice after cardiac arrest and cardiopulmonary resuscitation. IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 are immune inflammatory factors produced after activation of the STING signaling pathway, which can aggravate pathological damage.

[0041] Figure 4 The 7-day survival of mice in the STING inhibitor H151 administration group (15 mg / kg) and the solvent group after cardiac arrest and cardiopulmonary resuscitation ( Figure 4 A), neurological function scores at 3, 7, and 14 days ( Figure 4 B) and representative histological HE staining images of brain tissue ( Figure 4 C);

[0042] Figure 5The RT-qPCR experiment provided in the examples of the present invention detected the expression of IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 in brain tissue of mice in the sham group, the solvent group, and the STING inhibitor H151 administration group (15 mg / kg) after cardiac arrest and cardiopulmonary resuscitation. IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 are immune inflammatory factors produced after activation of the STING signaling pathway and can aggravate pathological damage;

[0043] Figure 6 The changes in the phosphorylation levels of IRF3 and TBK1, key proteins in the STING downstream signaling pathway, in the sham group, solvent group, and STING inhibitor H151 administration group (15 mg / kg) of mice after cardiac arrest and cardiopulmonary resuscitation provided in the embodiments of the present invention are shown. The upregulation of the phosphorylation levels of IRF3 and TBK1 indicates activation of the STING signaling pathway.

[0044] Figure 7 The expression of tight junction proteins ZO-1, Claudin-5, and Occludin in brain tissues of mice in the sham group, solvent group, and STING inhibitor H151 administration group (15 mg / kg) after cardiac arrest and cardiopulmonary resuscitation provided in the embodiments of the present invention was detected by immunoblotting and immunofluorescence experiments. The levels of tight junction proteins can reflect changes in blood-brain barrier permeability. When the expression level of tight junction proteins is downregulated, it indicates that the blood-brain barrier permeability is increased and the blood-brain barrier damage is aggravated;

[0045] Figure 8 Representative trajectory diagrams of the open field test of mice in the ham group, solvent group, and STING inhibitor H151 administration group (15 mg / kg) after cardiac arrest and cardiopulmonary resuscitation provided in the embodiments of the present invention ( Figure 8 A) Time of traveling to the central area ( Figure 8 B) and mileage and drop time in the rotating rod fatigue test ( Figure 8 C) Analysis. DETAILED DESCRIPTION

[0046] The technical scheme of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and specific embodiments, but it will be understood by those skilled in the art that the following described embodiments are part of embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. Those who do not specify specific conditions in the embodiments are carried out according to normal conditions or the conditions recommended by the manufacturer. Those whose reagents or instruments are not specified by the manufacturer are conventional products that can be purchased commercially.

[0047] The STING inhibitor (H-151) was provided by MCE (China) with the CAS number 941987-60-6. Six-week-old C57BL / 6J mice and STING knockout C57BL / 6J mice were purchased from the Institute of Animal Models, Nanjing University, and housed at the Animal Experimental Center, West China Hospital, Sichuan University. All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals and approved by the Experimental Animal Ethics Committee of West China Hospital, Sichuan University. Antibodies were purchased from Cell Signaling Technology, Inc., USA.

[0048] Example 1 Construction of a mouse model of post-cardiac arrest syndrome

[0049] (1) Preparation before inducing tremor

[0050] ① Anesthesia: First, induce anesthesia with 5% isoflurane, then continuously introduce pure oxygen through a mask, and maintain anesthesia with 1.5% isoflurane. Clamp the mouse's neck skin with toothed forceps. After there is no pain stimulation response, fix the mouse in a supine position on the operating table.

[0051] ② Endotracheal intubation: Use ophthalmic forceps in your left hand to penetrate the mouse's oropharynx and lift the soft palate to fully expose the glottis. With your right hand, insert a modified 20G intravenous indwelling endotracheal tube into the trachea along the glottis. After passing through the glottis, remove the guidewire and continue to insert the tube. The intubation depth is 2 cm from the front end of the endotracheal tube to the mouse's incisors.

[0052] ③ Right external jugular vein cannulation: Shave the skin on the right anterior neck of the mouse and disinfect it with iodine. Using ophthalmic forceps in the left hand, lift the skin and make a longitudinal incision 1 cm to the right of the midline of the mouse's neck. Separate the subcutaneous tissue layer by layer to fully expose the external jugular vein. Ligate the distal end with No. 7 silk suture. Puncture the vessel from the distal end to the proximal end with a 24G intravenous cannula. Advance the needle parallel to the vein 1-2 mm before withdrawing the cannula. Once blood returns, continue to advance the cannula proximally for approximately 3-5 mm before withdrawing the cannula. Secure the cannula with silk suture and seal the cannula with a heparin cap.

[0053] ④ Transesophageal fibrillation electrode insertion: Lubricate the esophageal fibrillation electrode with paraffin oil. Lift the mouse's tongue with curved forceps in your right hand. Slowly insert the esophageal electrode with your left hand. Adjust the electrode insertion depth based on the mouse's size, typically about 3-4 cm, ensuring the electrode rests against the posterior wall of the heart. Observe the ECG waveform to confirm electrode placement. Insert copper needle electrodes subcutaneously into the right upper, right, and left lower limbs of the mouse. Connect the electrodes to the BIOPAC system MP160 physiological signal acquisition system and record the ECG. Insert the copper electrode needle approximately 3-4 mm subcutaneously. Connect the fibrillation electrode and subcutaneous electrode needle to the BIOPAC system MP160 programmable stimulator.

[0054] (2) Establishment of post-cardiac arrest syndrome model

[0055] ①BIOPAC system MP160 stimulator program-controlled stimulation:

[0056] Discontinue anesthesia, disconnect the ventilator, and activate the BIOPAC system MP160 stimulator. Stimulate continuously for 120 seconds using the following parameters: output voltage 5V, output frequency 100Hz, and pulse width 10ms. Observe for 1-2 seconds after stopping stimulation. Successful induction of fibrillation is determined when the mouse exhibits respiratory arrest or cyanosis of the lips. When the ECG monitor indicates restoration of sinus rhythm, stimulate again for 30 seconds until sustained ventricular fibrillation, pulseless electrical activity, or cardiac arrest is induced.

[0057] ② Cardiopulmonary resuscitation based on small animal chest compression device:

[0058] When mice experienced persistent ventricular fibrillation, pulseless electrical activity, or cardiac arrest for 5 minutes, chest compressions were performed using a self-developed small animal chest compression device (Patent No. 201811350405.2). Parameters were: compression depth: 4 mm, compression rate: 300 times / min, and compression waveform: trapezoidal. At the start of cardiopulmonary resuscitation, mechanical ventilation was initiated with 100% oxygen concentration, 100 respiratory rate / min, tidal volume 12 ml / kg, and an inspiration-to-expiration ratio of 1:1. During cardiopulmonary resuscitation, 0.1 ml of epinephrine at a concentration of 50 g / ml was injected via the jugular vein, repeated every 3 minutes. ECG monitoring was monitored, and chest compressions were discontinued when spontaneous circulation returned. A heating pad was used to maintain the mouse's body temperature at 32.0 ± 0.2°C. When the mouse achieved recovery of spontaneous circulation (ROSC) for 5 min, FiO2 was adjusted to 50%. When the respiratory rate reached 60 breaths / min with a regular rhythm, the ventilator was stopped and the endotracheal tube was removed.

[0059] (3) Cardiac arrest, return of spontaneous circulation, and trial exit criteria

[0060] ① Cardiac arrest criteria:

[0061] ECG monitoring indicated pulseless electrical activity, ventricular fibrillation or cardiac arrest; absence of cardiac beats; and obvious cyanosis of the skin and mucous membranes of the lips, limbs, and tail.

[0062] ② Rest of spontaneous circulation criteria:

[0063] ECG monitoring showed supraventricular rhythm (including sinus, atrial or junctional rhythm); heart rate>200 beats / min, breathing recovery, physiological reflex recovery; obvious apical cardiac beats were visible; cyanosis of the mouse skin and mucous membranes was alleviated.

[0064] ③Experimental exit criteria:

[0065] The mice suffered cardiac arrest before esophageal electrode induction; cardiopulmonary resuscitation continued for 10 minutes without recovery of spontaneous circulation; and the mice suffered cardiac arrest again after recovery of spontaneous circulation.

[0066] Example 2

[0067] STING knockout mice (CPR-STING - / - ), 7-day survival of wild-type mice after cardiac arrest and cardiopulmonary resuscitation, neurological function scores on days 3, 7, and 14, and representative histological HE staining of brain tissue to evaluate brain tissue edema. Figure 1 shown.

[0068] Thirty wild-type C57BL / 6J mice were randomly divided into Sham group, CPR-WT group and CPR-STING group using random number table method. - / - The sham group of mice underwent anesthesia, endotracheal intubation, external jugular vein cannulation, and electrocardiogram (ECG) monitoring. After the mice regained consciousness, the endotracheal and external jugular vein cannulation were removed, the skin was sutured, and the mice were returned to their cages. In the experimental groups of wild-type mice and STING-knockout mice, the mice underwent anesthesia, endotracheal intubation, external jugular vein cannulation, and ECG monitoring to establish a post-cardiac arrest syndrome model. After the mice regained consciousness, the endotracheal and external jugular vein cannulation were removed, the skin was sutured, and the mice were returned to their cages.

[0069] Mice returned to their cages after ROSC were observed for 7 consecutive days, and their survival was recorded. Survival curves were drawn to calculate the survival rate of mice in each group 7 days after ROSC. Observers assessed the neurological function of mice daily after ROSC using the Neurological Function Score (NFS) chart and recorded it in detail.

[0070] Survival analysis results showed that compared with the CPR-WT group, the CPR-STING - / - The survival rate of mice in the CPR-STING group was significantly improved, with statistical significance. - / -The NFS of mice in each group was significantly increased, suggesting that knocking out the STING gene can alleviate brain damage after resuscitation and is beneficial to neurological prognosis.

[0071] HE staining results showed that the cerebral cortex of mice in the sham group had clear structure, cells were neatly arranged, and the density was uniform. The cell nuclei were light blue, round or oval, and the size and shape were normal. - / - The brain tissue damage in the CPR group was significantly reduced compared with that in the CPR-WT group, further suggesting that knocking out STING may alleviate brain tissue damage after resuscitation.

[0072] Example 3

[0073] Detection of Sham group, CPR-WT group, CPR-STING - / - The changes in the phosphorylation levels of IRF3 and TBK1, key proteins in the STING downstream signaling pathway, in the mice group after cardiac arrest and cardiopulmonary resuscitation were as follows. Figure 2 shown.

[0074] After the modeling was completed, the mice were killed by cervical dislocation under isoflurane anesthesia, and the brain tissue was removed. 20 mg of brain tissue was accurately weighed to extract protein to detect the expression changes of the phosphorylated versions of IRF3 and TBK1, key proteins in the STING downstream signaling pathway in the brain tissue.

[0075] The experimental results showed that compared with the wild-type mouse group (CPR-WT), CPR-STING - / - The protein expressions of p-IRF3 and p-TBK1 in the brain tissue of the mice in the control group were significantly downregulated, suggesting that the STING signaling pathway and its downstream immune inflammatory pathways such as type I interferon were blocked.

[0076] Example 4

[0077] RT-qPCR experiments detected the expression of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10 and CCL5 in the Sham group, CPR-WT group and CPR-STING group. - / - The expression of brain tissue in mice after cardiac arrest and cardiopulmonary resuscitation was shown in the following table. Figure 3 shown.

[0078] After modeling, mice were killed by cervical dislocation under isoflurane anesthesia. Brain tissue was removed, and 20 mg of brain tissue was accurately weighed for RNA extraction and testing. RT-qPCR reaction systems were prepared using TaKaRa SYBR Premix ExTaq reagent in a 20 μl reaction volume. Each sample was run in triplicate on a QuantStudio3 (Thermo Fisher Scientific) PCR instrument in a 96-well plate. Ct values ​​were collected using a calibrated threshold, and relative quantification was performed using real-time fluorescence quantitative PCR using β-actin as an internal reference gene. Expression of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 was detected.

[0079] The experimental results showed that compared with the wild-type mouse group (CPR-WT), CPR-STING - / - The gene expressions of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10 and CCL5 in the brain tissue of the mice in the STING group were significantly downregulated, suggesting that knocking out STING can reduce the activation of immune inflammatory pathways such as type I interferon.

[0080] Example 5

[0081] The 7-day survival of mice in the STING inhibitor H151 administration group and the solvent group after cardiac arrest and cardiopulmonary resuscitation, the neurological function scores on days 3, 7, and 14, and the representative histological HE staining of brain tissue and the evaluation of brain tissue edema were analyzed. Figure 4 shown.

[0082] Forty-five wild-type C57BL / 6J mice were randomly divided into a sham group, a CPR-vehicle group, and an H151-treated group (15 mg / kg) using a random number table, with n = 15 per group. A post-cardiac arrest syndrome model was established. Mice in the sham group underwent anesthesia, endotracheal intubation, external jugular vein cannulation, and electrocardiogram (ECG) monitoring. After the mice regained consciousness, the endotracheal and jugular vein cannulation were removed, the skin was sutured, and the mice were returned to their cages. In the wild-type mice and STING knockout mice groups, the mice underwent anesthesia, endotracheal intubation, external jugular vein cannulation, and ECG monitoring to establish a CA-CPR-ROSC model. After the mice regained consciousness, the endotracheal and jugular vein cannulation were removed, the skin was sutured, and the mice were returned to their cages. Mice returned to their cages after ROSC were observed for 7 consecutive days, and their survival was recorded. Survival curves were drawn to calculate the survival rate of mice in each group 7 days after ROSC.

[0083] The observers evaluated the neurological function of mice after ROSC every day according to the neurological function score (NFS) table and recorded it in detail. The results showed that the 7-day survival rate of the STING inhibitor H151 administration group (H151-CPR group) was higher than that of the solvent group (vehicle-CPR group). At the same time, the neurological function scores of the H151-CPR group on the 3rd and 7th days were also higher than those of the vehicle-CPR group, indicating that inhibiting the activation of the STING signaling pathway can improve the 7-day survival rate and neurological function of mice after CA-CPR-ROSC. HE staining results showed that CPR-STING - / - The brain tissue damage in the CPR group was significantly alleviated compared with that in the CPR-WT group.

[0084] Example 6

[0085] The expression of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10 and CCL5 in the brain tissue of mice in the Sham group, solvent group and H151 administration group (15 mg / kg) after cardiac arrest and cardiopulmonary resuscitation was detected. Figure 5 shown.

[0086] After modeling, mice were killed by cervical dislocation under isoflurane anesthesia. Brain tissue was removed, and 20 mg of brain tissue was accurately weighed for RNA extraction and testing. RT-qPCR reaction systems were prepared using TaKaRa SYBR Premix ExTaq reagent in a 20 μl reaction volume. Each sample was run in triplicate on a QuantStudio3 (Thermo Fisher Scientific) PCR instrument in a 96-well plate. Ct values ​​were collected using a calibrated threshold, and relative quantification was performed using real-time fluorescence quantitative PCR using β-actin as an internal reference gene. Expression of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10, and CCL5 was detected.

[0087] The experimental results showed that compared with the wild-type mouse group (CPR-WT), the gene expression of immune inflammatory molecules such as IFN-β, IL-1β, IL-6, TNF-α, CXCL10 and CCL5 in the brain tissue of mice in the H151 treatment group (15 mg / kg) was significantly downregulated, suggesting that knocking out STING can reduce the activation of immune inflammatory pathways such as type I interferon.

[0088] Example 7

[0089] The changes in the phosphorylation levels of IRF3 and TBK1, key proteins in the STING downstream signaling pathway, were detected in the sham group, solvent group, and H151-treated mice after cardiac arrest and cardiopulmonary resuscitation. Figure 6 shown.

[0090] After the modeling was completed, the mice were killed by cervical dislocation under isoflurane anesthesia, and the brain tissue was removed. 20 mg of brain tissue was accurately weighed to extract protein to detect the expression changes of the phosphorylated versions of IRF3 and TBK1, key proteins in the STING downstream signaling pathway in the brain tissue.

[0091] The experimental results showed that compared with the solvent group, the protein expression of p-IRF3 and p-TBK1 in the brain tissue of mice in the H151 treatment group (15 mg / kg) was significantly downregulated, suggesting that the STING signaling pathway and its downstream immune inflammatory pathways such as type I interferon were blocked.

[0092] Example 8

[0093] The blood-brain barrier damage in the brain tissue of mice in the sham group, solvent group, and H151-treated group after cardiac arrest and cardiopulmonary resuscitation was detected by immunoblotting and immunofluorescence experiments. Figure 7 shown.

[0094] After the modeling was completed, the mice were killed by cervical dislocation under isoflurane anesthesia, the brain tissue was removed, and 20 mg of brain tissue was accurately weighed to extract protein to detect the expression changes of tight junction proteins related to the blood-brain barrier in the brain tissue.

[0095] The experimental results showed that compared with the solvent group, the expression of tight junction proteins ZO-1, Claudin-5, and Occludin in the brain tissue of mice in the H151 treatment group (15 mg / kg) was significantly upregulated, suggesting that the STING inhibitor H151 can significantly reduce the blood-brain barrier damage in mice with post-cardiac arrest syndrome. Immunofluorescence staining was used to detect the expression levels of Occludin, Claudin-5, and ZO-1 proteins in cerebral microvascular endothelial cells in the cerebral cortex, which also confirmed that the STING inhibitor H151 can upregulate the level of tight junction proteins in the blood-brain barrier of mice with post-cardiac arrest syndrome and reduce blood-brain barrier damage.

[0096] Example 9

[0097] The behavioral abilities of mice in the sham group, solvent group, and H151 administration group (15 mg / kg) after cardiac arrest and cardiopulmonary resuscitation were tested. Figure 8 shown.

[0098] Forty-five wild-type C57BL / 6J mice were randomly divided into a sham group, a CPR-vehicle group, and an H151-treated group (15 mg / kg) using a random number table, with n = 15 per group. Behavioral training was performed one day before modeling to obtain baseline data. A post-cardiac arrest syndrome model was established. Rotarod fatigue tests were performed on days 3, 7, and 14 after modeling, and an open field test was performed on day 7, and data were collected.

[0099] The results showed that the H151-treated group increased the time and distance traveled in the central area compared to the vehicle group, but had no effect on the total distance traveled. In the rotarod fatigue test, the H151-treated group significantly increased the time spent exploring the novel arm compared to the vehicle group 7 and 14 days after modeling. Therefore, the STING inhibitor H151 can significantly improve the spatial memory and motor function of mice with post-cardiac arrest syndrome.

[0100] In summary, knocking out the STING gene or using STING inhibitors can significantly reduce brain tissue damage in mice with post-cardiac arrest syndrome by regulating the STING-I type interferon immune-inflammatory signaling pathway, improve the spatial memory and motor functions of mice, and significantly increase the overall survival and survival rate of mice with post-cardiac arrest syndrome.

[0101] Although the present invention has been illustrated and described using specific embodiments, it should be appreciated that the above embodiments are merely intended to illustrate the technical solutions of the present invention rather than to limit them. Those skilled in the art should understand that the technical solutions described in the above embodiments may be modified, or some or all of the technical features thereof may be replaced by equivalents, without departing from the spirit and scope of the present invention. However, these modifications or replacements do not deviate from the essence of the corresponding technical solutions within the scope of the technical solutions of the embodiments of the present invention. Therefore, this means that all such replacements and modifications within the scope of the present invention are included in the appended claims.

Claims

1. Use of STING inhibitors in the preparation of drugs for the treatment and / or prevention of post-cardiac arrest syndrome; The STING inhibitors include: H-151； The application includes any one or more of the following: (1) Alleviate brain damage caused by post-cardiac arrest syndrome; (2) Improve spatial memory function and / or motor function after post-cardiac arrest syndrome; (3) Improve the survival period and survival rate after post-cardiac arrest syndrome.

2. The use according to claim 1, characterized in that The STING inhibitor can regulate the STING-mediated type I interferon immune inflammatory signaling pathway.

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